Selective inhibitors of the urokinase plasminogen activator

ABSTRACT

The present invention relates to novel selective inhibitors of the urokinase plasminogene activator (uPA, EC 3.4.21.31) of the aryl guanidine type.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a National Stage entry of International ApplicationNo. PCT/EP00/08234, filed Aug. 23, 2000, the entire specification claimsand drawings of which are incorporated herewith by reference.

The present invention relates to novel selective inhibitors of theurokinase plasminogen activator (uPA, EC 3.4.21.31) of the arylguanidinetype.

The urokinase-type plasminogen activator (uPA) plays a key part in tumorinvasion and formation of metastases (Schmitt et al., J. Obst. Gyn. 21(1995), 151–165). uPA is overexpressed in various types of tumor cells(Kwaan, Cancer Metastasis Rev. 11 (1992), 291–311) and binds to thetumor-associated uPA receptor (uPA-R) in which activation of plasminogento plasmin takes place. Plasmin is capable of degrading variouscomponents of the extracellular matrix (ECM) such as fibronectin,laminin and collagen type IV. It also activates some other ECM-degradingenzymes, in particular matrix metalloproteinases. High amounts oftumor-associated uPA correlate with a higher risk of metastasizing incancer patients (Stephens et al., Breast Cancer Res. & Treat. 52 (1998),99–111). Therefore, inhibition of the proteolytic activity of uPA is agood starting point for an anti-metastatic therapy.

A common feature of many known synthetic uPA inhibitors is a basicresidue containing amidino or guanidino groups, which can bind to Asp¹⁸⁹in the uPA S1 specificity pocket and which acts as an arginine mimeticthere (Spraggon et al., Structure 3 (1995), 681–691). However, most ofthe known inhibitors are not selective for uPA but also inhibit otherserine proteases such as trypsin, thrombin, plasmin or tissueplasminogen activator (tPA).

p-Aminobenzamidine is a moderately selective uPA inhibitor having aninhibition constant of 82 μM. Billstroem et al. (Int. J. Cancer 61(1995); 542–547) could show a distinct decrease in the growth rate ofDU145 tumors (a prostate adenocarcinoma cell line) in SCID mice whenadministering orally a daily dose of 125 to 250 mg ofp-aminobenzamidine/kg/day. The side effects were negligible.

Some monosubstituted phenylguanidines have proved effective andselective uPA inhibitors in vitro. These small molecules show inhibitionconstants in the micromolar range but they bind only in the S1 pocket ofuPA (Yang et al., J. Med. Chem. 33 (1990), 2956–2961). Biologicalstudies using these compounds were not carried out.

The diuretic amiloride is a selective uPA inhibitor (Ki, uPA=7 μM) whichprevents the formation of lung metastases after i.v. inoculation of ratbreast adenocarcinoma cells (Kellen et al., Anticancer Res. 8 (1988),1373–1376). Some 3-amidinophenylalanine derivatives have likewise provedeffective inhibitors of serine proteases but these compounds generallyhave only low selectivity for uPA (Stürzebecher et al., J. Med. Chem. 40(1997), 3091–3099; Stürzebecher et al., J. Enzyme Inhib. 9 (1995),87–99).

Currently the most effective and most selective uPA inhibitors arebenzo[b]thiophene-2-carboxamidine derivatives (B428 and B623:K_(i′ uPA)=0.32 and 0.07 μM, respectively; U.S. Pat. No. 5,340,833).Rabbani et al. (Int. J. Cancer 63 (1995), 840–845) and also Xing et al.(Cancer Res. 57 (1997), 3585–3593) could show, after administration of4-iodobenzo[b]thiophene-2-carboxamidine (B428), a decrease of tumorgrowth and metastases formation in a syngeneic model of rat prostatecancer and mouse breast cancer, respectively. The latter studies showeda further decrease in primary tumor growth when B428 was administeredtogether with the antiestrogen tamoxifen.

It was the object of the present invention to provide novel selectiveuPA inhibitors. This object is achieved by novel arylguanidine and inparticular phenylguanidine derivatives. These compounds contain afurther substituent on the aromatic ring system, preferably in paraposition to the guanidine group, which substituent contains anunsubstituted or substituted methylene group followed by hydrogendonor/acceptor functionalities. Owing to this substitution pattern, thecompounds are particularly effective and selective for uPA. Thisefficacy could be attributed possibly to the fact that they

-   (1) interact as arginine mimetics with the Asp¹⁸⁹ amino acid residue    in the S1 pocket of uPA and-   (2) can interact with the S2 and/or S3 pockets of uPA.

N-Substituted p-aminophenylguanidines (without methylene spacer) andalso p-guanidinophenylalanine derivatives (2 methylene groups as spacer)were ineffective uPA inhibitors. The compounds of the inventionpreferably contain urethane or urea groups for interaction with S2and/or large hydrophobic radicals such as aryl groups or cycloalkylgroups (e.g. adamantane) for interaction with S3.

The present invention thus relates to the use of compounds of theformula I

in which

-   -   Ar is an aromatic or heteroaromatic ring system,    -   X¹ is NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵,    -   R¹ is H, an unsubstituted or substituted alkyl, alkenyl,        alkynyl, aryl or/and heteroaryl radical, or COOR³, CONR³R⁴ or        COR⁵,    -   R² is halogen, C(R⁶)₃), C₂(R⁶)₅, CO(R⁶)₃ or OC₂(R⁶)₅,    -   R³ is H or any organic radical,    -   R⁴ is H or an unsubstituted or substituted alkyl, alkenyl or        alkynyl radical,    -   R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl,        carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl        radical, where the alkyl, alkenyl, alkynyl, aryl and heteroaryl        radicals may be unsubstituted or substituted,    -   R⁶ is in each case independently H or halogen, in particular F,        and    -   m is an integer from 0 to 4,    -   or salts of said compounds for preparing an agent for inhibition        of the urokinase plasminogen activator.

The compounds may be present as salts, preferably as physiologicallytolerated acid salts, for example as salts of mineral acids,particularly preferably as hydrochlorides or as salts of suitableorganic acids. The guanidinium group may carry, where appropriate,protective functions which are removable by cleavage, preferably underphysiological conditions. The compounds may be present as optically purecompounds or as mixtures of enantiomers or/and diastereoisomers.

In the compounds of the general formula (I), Ar is preferably anaromatic or heteroaromatic ring system having a single ring, inparticular a benzene ring. In this ring system the substituents CHX¹R¹and NHC(NH)NH₂ are preferably arranged in meta or para position andparticularly preferably in para position. In addition, Ar may furthercontain other, non-hydrogen substituents R². The number of substituentsR² is preferably 0, 1, 2 or 3, particularly preferably 0 or 1 and mostpreferably 0. Preferred examples of R² are halogen atoms (F, Cl, Br orI), CH₃, CF₃, OH, OCH₃ or OCF₃.

The substituent —CHX¹R¹ is critical for inhibitor activity. R¹ may be Hor an unsubstituted or substituted alkyl, alkenyl, alkynyl, aryl or/andheteroaryl radical. The alkyl radical may be a straight-chain orbranched C₁–C₁₀-alkyl group, in particular a C₁–C₄-alkyl group or aC₃–C₈-cycloalkyl group which may be substituted with, for example,C₁–C₃-alkoxy, hydroxyl, carboxyl, amino, sulfonyl, nitro, cyano, oxoor/and halogen or else with aryl or heteroaryl radicals. Alkenyl andalkynyl radicals are preferably C₂–C₁₀ groups, in particular C₂–C₄groups which may be unsubstituted or substituted as described above.Aryl and heteroaryl radicals may be substituted, for example, withC₁–C₆-alkyl, C₁–C₃-alkoxy, hydroxyl, carboxyl, sulfonyl, nitro, cyanoor/and oxo. Furthermore, R¹ may have the meanings COOR³, CONR³R⁴ orCOR⁵.

The X¹ group is a radical having electron donor or/and electron acceptorproperties, preferably NR³R⁴, OR³, SR³ COOR³, CONR³R⁴ or COR⁵. X¹ isparticularly preferably NR³R⁴. R³ may be any organic radical orhydrogen. R⁴ may be hydrogen or an unsubstituted or substituted alkyl,alkenyl or alkynyl radical, as described above.

R⁵ may be hydrogen or an alkyl, alkenyl, alkynyl, carboxyalkyl,carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroarylradical. R⁵ is preferably a space-filling radical and contains at leastone aryl, heteroaryl, cycloalkyl or/and tert-alkyl group. Particularpreference is given to phenyl radicals, substituted phenyl radicals,tert-alkyl radicals and cycloalkyl radicals, which may contain, whereappropriate, substituents as defined above.

If X¹ has the meaning NR³R⁴ and R³ and R⁴ are in each case independentlyhydrogen or unsubstituted or substituted alkyl, alkenyl, alkynyl orheteroaryl radicals (see definition of R¹), R¹ has preferably a meaningdifferent from hydrogen, particularly preferably COOR³, CONR³R⁴ or COR⁵,in particular COOR³, CONH₂, CO—COOR⁵ or CHO so that the compounds I arederivatives of guanidinophenylglycine.

R³ is particularly preferably a group of the general formula (II):

in which

-   X² is NH, NR⁴, O or S,-   X³ is NH, NR⁴, O, S, CO, COO, CONH or CONR⁴,-   Y is C(R⁸)₂,-   R⁴ is defined as in formula (I),-   R⁷ is H or an unsubstituted or substituted alkyl, alkenyl, alkynyl,    aryl or/and heteroaryl radical or —SO₂—R⁹,-   R⁸ is in each case independently H, halogen or an unsubstituted or    substituted alkyl, alkenyl, alkynyl or aryl or/and heteroaryl    radical,-   R⁹ is H or an unsubstituted or substituted alkyl, alkenyl, alkynyl,    aryl or/and heteroaryl radical and-   n is an integer from 0 to 2.

X² is preferably NH or O, particularly preferably O. X³ is preferably NHor —O—. Y is preferably CH₂ or CHR⁸, R⁸ being preferably defined as R⁴in formula (I).

R⁷ and R⁹ are preferably defined as R⁵ in formula (I).

R³ is most preferably a group of the formula IIIa, IIIb or IIIc:

in which R⁷ and R⁹ are as defined in formula (II).

The substituents R⁷ and R⁹ contain, like R⁵, preferably space-fillinggroups which may be selected from the group comprising unsubstituted orsubstituted aryl radicals, in particular phenyl and substituted phenylradicals and unsubstituted or substituted branched alkyl, alkenyl oralkynyl radicals, in particular with tertiary carbon atoms such astert-butyl or neopentyl, or unsubstituted or substituted cycloalkylradicals, in particular bi- or tricycloalkyl radicals such as adamantyl.

Particularly high affinity and selectivity for uPA are also exhibited bycompounds of the general formula (IV):

-   in which Ar, X¹, R² and m, on each occurrence, independently may be    identical or different and have a meaning as defined in the formulae    (I), (II) and (IIIa–c).

The compounds of the formula (IV) contain two arylguanidino groups andare linked to one another via their substituents CHR¹X¹— which may be ineach case identical or different.

The compounds of the general formula (I) may be prepared, for example,starting from p-aminobenzylamine according to the reaction schemes shownin FIGS. 1 and 2. For example, 4-aminobenzylamine may be reacted with aprotective reagent for amino groups, for example di-tert-butylpyrocarbonate, to give a protected intermediate,4-(N-Boc-aminomethyl)aniline (1), Boc meaning tert-butyloxycarbonyl. Thearomatic amino function of this compound can be reacted with aguanidinylation reagent, for example N,N′-di-Z-N″-triflylguanidine,resulting in 1-[4-(N-Boc-aminomethyl)phenyl-2,3-di-Z-guanidine (2), Zbeing benzyloxycarbonyl. This compound can be converted to1-[4-(aminomethyl)phenyl]-2,3-di-Z-guanidinium hydrochloride (4) byremoving the Boc protective group by cleavage. The compound (4) may inturn be reacted with reactive compounds such as, for example,chloroformic esters, isocyanates or N-hydroxysuccinimide esters to givethe desired final products.

The preparation of hydrogenation-labile compounds is described in FIG.2. 4-Aminobenzylamine can be reacted with a protective reagent for aminogroups, for example benzyloxycarbonyloxysuccinimide to give a protectedintermediate (6) and then with a further guanidinylation reagent, forexample N,N′-di-Boc-1-guanylpyrazole, to give (7). This compound can behydrogenated to give (8) and then be reacted with reactive compounds togive the desired final products.

Correspondingly, it is also possible to synthesize compounds in which X¹has the meaning OR³, SR³, COOR³, CONR³R⁴ or COR⁵.

The urokinase inhibitors of the invention may be used, whereappropriate, together with suitable pharmaceutical auxiliary agents orcarriers for producing medicaments or in diagnostics. In thisconnection, administration in combination with other active substances,for example other urokinase inhibitors such as, for example, antibodiesor/and peptides, is possible.

The medicaments may be administered in humans and animals topically,orally, rectally or parenterally, for example subcutaneously orintravenously, for example in the form of tablets, coated tablets,capsules, pellets, suppositories, solutions or transdermal systems suchas plasters.

The compounds of the invention are suitable for controlling disorderswhich are associated with pathological overexpression of uPA or/anduPAR. They are, for example, capable of very effectively inhibiting thegrowth or/and spreading of malignant tumors and also metastasizing oftumors. It is possible to use the uPA inhibitors, where appropriate,together with other tumor agents or with other types of treatment, forexample radiation or surgery. Furthermore, the inhibitors of theinvention are also effective in other uPA-associated disorders.

uPA inhibitors of the invention are preferably characterized in thatthey have a K_(i) which is at least two times, preferably at least fivetimes and particularly preferably at least ten times and up to 1 000times lower for uPA than for tPA. It is furthermore remarkable that thecompounds of the invention only marginally affect blood clotting, sincetheir K_(i) values are too high for effective inhibition of thrombin,plasmin and factor Xa.

The inventive substances of the formula (I) may be used in the form ofconjugates with physiologically effective substances, for exampleradiolabels or cytotoxic agents, e.g. chemotherapeutics such ascisplatin or 5-fluorouracil, or with peptides. Furthermore, it is alsopossible to incorporate the substances into the membrane of carriervesicles, for example liposomes, and thus to make possible targeting ofactive substances enclosed in said carrier vesicles, for examplecytotoxic agents such as doxorubicin.

The present invention provides a method for inhibiting urokinase inliving creatures, in particular in humans, by administering an effectivequantity of at least one compound of the formula (I). The dosage of thecompound is commonly in the range from 0.01 to 100 mg/kg of body weightper day. The length of treatment depends on the seriousness of thedisorder and may range from a single dose up to a treatment lastingseveral weeks or even several months, which may be repeated atintervals, where appropriate.

Finally, the present invention relates to novel arylguanidinederivatives of the general formula (I).

The invention is intended to be illustrated in more detail by thefollowing examples and figures in which:

FIG. 1 shows a general reaction scheme for preparinghydrogenation-stable substances of the invention, and

FIG. 2 shows a general reaction scheme for preparinghydrogenation-labile substances of the invention.

EXAMPLES

Materials and Methods

All solvents and reagents used for the synthesis of uPA inhibitors wereof the highest commercially available quality and were, if necessary,further purified and dried by standard methods. Analytical HPLC wascarried out on Nucleosil 100/C18 columns (Macherey-Nagel, Düren,Germany) using a linear acetonitrile/2% H₃PO₄ gradient (from 5:95 to90:10 in 13 min). ESI-MS spectra were measured in a Perkin Elmer API 165mass spectrometer.

Example 1 Synthesis of Acid-Labile Urethanes, for Example4-(N-Boc-aminomethyl)phenylguanidine (3) 4-(N-Boc-Aminomethyl)aniline(1)

4-Aminobenzylamine (2 ml; 17.6 mmol) was dissolved in 1,4-dioxane (10ml). An aqueous 2 N NaOH solution (17.6 ml; 35.2 mmol) was added withstirring. A solution of di-tert-butyl pyrocarbonate (3.08 g; 14.1 mmol)in 1,4-dioxane (30 ml) was added dropwise over 30 min and the reactionmixture was stirred at room temperature overnight. The solution wasconcentrated under reduced pressure to approximately 10 ml and extractedtwice with ethyl acetate (30 ml). The combined organic phases werewashed with aqueous 5% KHSO₄ (10 ml), aqueous 5% NaHCO₃, water and saltsolution, dried over anhydrous Na₂SO₄ and evaporated under reducedpressure, the resulting product being a light yellow solid substance.

Yield: 2.38 g (76%); HPLC: t_(R) 5.6 min; MS 223 (M+H)⁺, calculated 222(M).

1-[4-(N-Boc-Aminomethyl)phenyl]-2,3-di-Z-guanidine (2)

A solution of the compound (1) (500 mg; 2.24 mmol) andN,N′-di-Z-N″-triflylguanidine (1.04 g; 2.24 mmol) (Feichtinger et al.,J. Org. Chem. 63 (1998), 38043805) in 5 ml of acetone was stirredvigorously at room temperature. After 10 min the product started toprecipitate. After 2 h the product was filtered off, dried under reducedpressure and recrystallized from methanol, resulting in white crystals.

Yield: 1.065 g (89%); HPLC: t_(R) 13.4 min; MS 533 (M+H)⁺, calculated532 (M).

4-(Boc-Aminomethyl)phenylguanidinium hydrochloride (3)

50 mg (0.107 mmol) of the compound (2) were dissolved in 5 ml ofmethanol, stirred and hydrogenated over a 10% palladium/activated carboncatalyst for 3 h. After removing the catalyst by filtration, the solventwas evaporated under reduced pressure. The residue was recrystallizedfrom methanol/diisopropyl ether after adding one equivalent of HCl in1,4-dioxane.

Yield: 28 mg (87%); HPLC: t_(R) 7.1 min; MS 265 (M+H)⁺, calculated 264(M).

Example 2 Synthesis of Disubstituted Ureas Using1-[4-(aminomethyl)phenyl]-2,3-di-Z-guanidinium hydrochloride (4) asComponent, for Example 4-[3-(1-adamantyl)ureido]-phenylguanidiniumhydrochloride (5) 1-[4-(Aminomethyl)phenyl]-2,3-di-Z-guanidiniumhydrochloride (4)

1 g (1.878 mmol) of the compound (2) was dissolved at 0° C. in 20 ml of3 N HCl (gas) in 1,4-dioxane and stirred at room temperature for 2 h.After evaporating the solvent, the crystalline product was obtained invirtually quantitative yield.

Yield: 872 mg (99%); HPLC: t_(R) 10.2 min; MS 433 (M+H)⁺, calculated 432(M).

4-[3-(1-adamantyl)ureido]phenylguanidinium hydrochloride (5)

50 mg (0.107 mmol) of the compound (4), 17 mg (0.107 mmol) of adamantylisocyanate and 45 μl (0.32 mmol) of triethylamine were dissolved in 1 mlof ethylene chloride. The reaction mixture was stirred at roomtemperature for 3 h. After evaporating the solvent under reducedpressure, the residue was dissolved in ethyl acetate (10 ml) andextracted three times with 0.1 N aqueous HCl. The organic phase wasconcentrated to dryness. The protective groups Z were removed asdescribed for compound (3).

Yield: 15 mg (37%); HPLC: t_(R) 8.6 min; MS 342 (M+H)⁺, calculated 341(M).

Example 3 Synthesis of Hydrogenation-Labile Compounds, for Example4-[N-(4-nitrobenzyloxycarbonyl)aminomethyl]phenylguanidine (9)4-(N-Z-Aminomethyl)aniline (6)

4-Aminobenzylamine (1 ml; 8.82 mmol) was dissolved in 10 ml of1,4-dioxane. An aqueous 2 N solution of NaOH (8.8 ml; 17.64 mmol) wasadded with stirring. Then a solution of benzyloxycarbonyloxysuccinimide(1.978 g; 7.938 mmol) in 10 ml of 1,4-dioxane was added dropwise over 15min, and the reaction mixture was stirred at room temperature for 5 h.The solution was concentrated under reduced pressure to approximately 10ml and extracted twice with 30 ml of ethyl acetate. The combined organicphases were washed with aqueous 5% strength NaHCO₃ solution, water andsalt solution, dried over anhydrous Na₂SO₄, concentrated and dried underreduced pressure, the resulting product being a light yellow solidsubstance.

Yield: 1.8 g (88%); HPLC: t_(R) 6.8 min; MS 257 (M+H)⁺, calculated 256(M).

1-[4-(N-Z-Aminomethyl)phenyl]-2,3-di-Boc-guanidine (7)

A solution of 495 mg (1.93 mmol) of the compound (6) and 599 mg (1.93mmol) of N,N′-di-Boc-1-guanylpyrazole (Bernatowicz et al., TetrahedronLett. 34 (1993), 3389–3392) in 5 ml of acetone was stirred at roomtemperature for 3 days. After evaporating the solvent, the residue wasdissolved in 50 ml of diethyl ether, washed with aqueous 5% KHSO₄solution, water and salt solution and dried over anhydrous Na₂SO₄.Evaporating the diethyl ether under reduced pressure resulted in a lightyellow foam.

Yield: 670 mg (70%); HPLC: t_(R) 12.1 min; MS 499 (M+H)⁺, calculated 498(M).

1-(4-Aminomethyl)phenyl-2,3-di-Boc-guanidine hydrochloride (8)

The compound (8) was obtained by catalytic hydrogenation of 600 mg (1.2mmol) of the compound (7) in ethanol over a 10% palladium/activatedcarbon catalyst for 1 h. After filtration of the catalyst, the solventwas evaporated under reduced pressure, resulting in an oil which wasrecrystallized from isopropanol/diisopropyl ether after adding 1equivalent of HCl in 1,4-dioxane.

Yield: 450 mg (91%); HPLC: t_(R) 8.1 min; MS 365 (M+H)⁺, calculated 364(M).

4-[N-(4-Nitrobenzyloxycarbonyl)aminomethyl]phenylguanidine hydrochloride(9)

A solution of 50 mg (0.125 mmol) of the compound (8), 27 mg (0.125 mmol)of 4-nitrobenzyl chloroformate and 52 μl (0.375 mmol) of triethylaminein 1 ml of methylene chloride was stirred at room temperature for 3 h.After evaporating the solvent, the residue was dissolved in 30 ml ofethyl acetate and washed three times with 0.5 N aqueous HCl. Afterevaporating the ethyl acetate, the residue was dissolved in 95%trifluoroacetic acid and stirred for 1 h. After evaporating the solvent,the product was recrystallized from ethanol/diisopropyl ether.

Yield: 35 mg (60%); HPLC: t_(R) 8.1 min; MS 344 (M+H)⁺, calculated 343(M).

Example 4 In-Vitro Inhibition of Urokinase by Selected Compounds of theFormula I

The uPA inhibitor activity was determined by incubating 200 μl of Trisbuffer (0.05 mol/l, containing the inhibitor, 0.154 mol/l NaCl, 5%ethanol, pH 8.0), 25 μl of substrate (Pefachrome UK orBZ-β-Ala-Gly-Arg-pNA in H₂O; Pentapharm Ltd, Basle, Switzerland) and 50μl of sc-urokinase (Ribosepharm GmbH, Haan, Germany) or anothercorresponding protease at 25° C. After 3 min, the reaction wasinterrupted by adding 25 μl of acetic acid (50%) and absorbance at 405nm was determined by means of a microplate reader (MR 5000, Dynatech,Denkendorf, Germany). The K_(i) values were determined by linearregression according to Dixon by means of a computer program. The K_(i)values are the average of at least three determinations, and thestandard deviation was below 25%. The inhibitors assayed and theirinhibition constants for various proteases are listed in table 1 below:

TABLE 1 Ki [μM] Inhibitor Name uPA Plasmin Thrombin Trypsin F Xa

ST 269 27 >1000 >1000 >1000 >1000

ST 270 46 >1000 >1000 >1000 >1000

ST 242 36 >1000 >1000 >1000 >1000

ST 274 13 >1000 >1000 >1000 >1000

ST 293 2,4 >1000 600 46 >1000

ST 282 240 >1000 >1000 >1000 >1000

ST 267 >1000 >1000 >1000 >1000 >1000

ST 296 22 >1000 >1000 42 >1000

ST 294 37 >1000 >1000 >1000 >1000

ST 298 42 >1000 >1000 37 >1000

ST 270 46 >1000 >1000 >1000 >1000

ST 271 51 >1000 >1000 >1000 >1000

ST 275 >1000 >1000 >1000 >1000 >1000

ST 273 52 130 >1000 >1000 >1000

ST 301 29 170 >1000 >1000 330

ST 311 12 ??? >1000 200 >1000

ST 312 2,8 ??? >1000 100 >1000

ST 313 35 ??? >1000 ??? >1000

ST 315 11 ??? >1000 200 >1000

The compounds ST293, 312, and 315 have a K_(i) value for uPA of >1 000μm.

The compounds denoted as ST293 and ST312 proved to be particularlyeffective and selective inhibitors.

1. A pharmaceutical composition comprising at least one compound of the formula (I)

in which the substituents —CH₂X¹ and —NHC(NH)NH₂ are arranged in a para position to each other; Ar is an aromatic or heteroaromatic ring system having a single ring; X¹ is NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is H or a group of the formula II, IIIa, IIIb or IIIc:

where X² is NH, NR⁴, O or S, X³ is NH, NR⁴, O, S, CO, COO, CONH OR CONR⁴, Y is C(R⁸)₂, R⁴ is H or an alkyl, alkenyl or alkynyl radical, R⁷ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical or —SO₂—R⁹, R⁸ is in each case independently H, halogen or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical, R⁹ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical and n is an integer from 0 to 2, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is halogen, C(R⁶)₃, C₂(R⁶)₅, OC(R⁶)₃ or OC²(R⁶)₅, where R⁶ is in each case independently H or halogen and m is an integer from 0 to 4; or a salts of said at least one compound, and a pharmaceutically acceptable carrier therefor.
 2. A pharmaceutical composition according to claim 1, in which Ar is a benzene ring.
 3. A pharmaceutical composition according to claim 1, in which R⁷ and R⁹ are each independently an aryl, tertiary alkyl or cycloalkyl radical.
 4. A pharmaceutical composition comprising at least one compound of the formula (IV)

in which X¹ is in each case independently NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is in each case independently H or any organic radical, R⁴ is in each case independently H or an alkyl, alkenyl or alkynyl radical; Ar is in each case independently an aromatic or heteroaromatic ring system, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is in each case independently halogen, C(R⁶)₃ ³, C₂(R⁶)₅, OC(R⁶)₃ or OC₂(R⁶)₅, where R⁶ is in each case independently H or halogen; and m is an integer from 0 to 4; or salts of said at least one compound, and a pharmaceutically acceptable carrier therefor.
 5. A pharmaceutical composition according to claim 1 wherein said composition is adapted to be administered orally, topically, rectally or parenterally.
 6. A pharmaceutical composition according to claim 1 wherein said composition is adapted to be administered in the form of tablets, coated tablets, capsules, pellets, suppositories, solutions or transdermal systems.
 7. A pharmaceutical composition according to claim 1, wherein said compound of the formula I has a K_(i) that is at least two times lower for uPA than for tPA.
 8. A pharmaceutical composition according to claim 1, wherein said compound of the formula I has a K_(i) that is at least five times lower for uPA than for tPA.
 9. A pharmaceutical composition according to claim 1, wherein said compound of the formula I has a K_(i) that is at least 10 times lower for uPA than for tPA.
 10. A pharmaceutical composition according to claim 1, wherein said compound of the formula I has a K_(i) that is at least 1000 times lower for uPA than for tPA.
 11. A pharmaceutical composition according to claim 1, wherein said compound of the formula I is conjugated with at least one physiological effective substance, wherein said substance is at least one radiolabelled substance.
 12. A pharmaceutical composition according to claim 1, wherein said compound of the formula I is incorporated into a carrier vesicle.
 13. A pharmaceutical composition according to claim 1, wherein R⁶ in said compound of formula I is F.
 14. A pharmaceutical composition according to claim 3, wherein said at least one aryl radical is a phenyl radical.
 15. A pharmaceutical composition according to claim 3, wherein said at least one cycloalkyl radical is a bicycloalkyl radical.
 16. A pharmaceutical composition according to claim 15, wherein said bicycloalkyl radical is an adamantyl radical.
 17. A pharmaceutical composition according to claim 4, wherein R⁶ in said compound of formula I is F.
 18. A compound of the formula (I)

in which the substituents —CH₂X¹ and —NHC(NH)NH₂ are arranged in a para position to each other; Ar is an aromatic or heteroaromatic ring system having a single ring; X¹ is NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is H or a group of the formula II, IIIa, IIIb or IIIc:

where X² is NH, NR⁴, O or S, X³ is NH, NR⁴, O, S, CO, COO, CONH OR CONR⁴, Y is C(R⁸)₂, R⁴ is H or an alkyl, alkenyl or alkynyl radical, R⁷ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical or —SO₂—R⁹, R⁸ is in each case independently H, halogen or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical, R⁹ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical and n is an integer from 0 to 2, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is halogen, C(R⁶)₃, C₂(R⁶)₅, OC(R⁶)₃ or OC²(R⁶)₅, where R⁶ is in each case independently H or halogen; and m is an integer from 0 to 4, with the provisos that when Ar=phenyl, m=O, CH₂X¹ is not CH₂COOH, when Ar=phenyl, m=0, X¹=NR³R⁴ with R⁴=H and R³=—COOR⁷ with R⁷=tertbutyl and m=0, the compound of formula (I) is not in the hydrochloride salt form, and when Ar=phenyl, m=0 and X¹=NH₂ the compound of formula (I) is not in the bistrifluoroacetate salt form.
 19. The compound of claim 18, in which Ar is a benzene ring.
 20. A compound of the formula (I)

in which Ar is an aromatic or heteroaromatic ring system having a single ring: X¹ is NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is H or a group of the formula II, IIIa, IIIb or IIIc:

where X² is NH, NR⁴, O or S, X³ is NH, NR⁴, O, S, CO, COO, CONH OR CONR⁴, Y is C(R⁸)₂, R⁴ is H or an alkyl, alkenyl or alkynyl radical, R⁷ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical or —SO₂—R⁹, R⁸ is in each case independently H, halogen or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical, R⁹ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical and n is an integer from 0 to 2, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is halogen, C(R⁶)₃, C₂(R⁶)₅, OC(R⁶)₃ or OC²(R⁶)_(5a) where R⁶ is in each case independently H or halogen; and m is an integer from 0 to 4, with the provisos that when Ar=phenyl, m=O, CH₂X¹ is not CH₂COOH, when Ar=phenyl, m=0, X¹=NR³R⁴ with R⁴=H and R³=—COOR⁷ with R⁷=tertbutyl and m=0, the compound of formula (I) is not in the hydrochloride salt form, and when Ar=phenyl, m=0 and X¹=NH₂ the compound of formula (I) is not in the bistrifluoroacetate salt form, in which R⁷ and R⁹ are at least one aryl radical, at least one tertiary alkyl radical or at least one cycloalkyl radical.
 21. A compound according to claim 20, wherein said at least one aryl radical is a phenyl radical.
 22. A compound according to claim 20, wherein said at least one cycloalkyl radical is a bicycloalkyl radical.
 23. A compound according to claim 22, wherein said bicycloalkyl radical is an adamantyl radical.
 24. A compound according to claim 18 wherein R⁶ in said compound of formula I is F.
 25. A compound of the formula (IV)

in which X¹ is in each case independently NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is in each case independently H or any organic radical, R⁴ is in each case independently H or an alkyl, alkenyl or alkynyl radical; Ar is in each case independently an aromatic or heteroaromatic ring system, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is in each case independently halogen, C(R⁶)₃ ³, C₂(R⁶)₅, OC(R⁶)₃ or OC₂(R⁶)₅, where R⁶ is in each case independently H or halogen; and m is an integer from 0 to 4; or salts of said compound.
 26. A pharmaceutical composition comprising at least one compound of the formula (I)

in which Ar is an aromatic or heteroaromatic ring system having a single ring; X¹ is NR³R⁴, OR³, SR³, COOR³, CONR³R⁴ or COR⁵, where R³ is a group of the formula II, IIIa, IIIb or IIIc:

where X² is NH, NR⁴, O or S, X³ is NH, NR⁴, O, S, CO, COO, CONH OR CONR⁴, Y is C(R⁸)₂, R⁴ is H or an alkyl, alkenyl or alkynyl radical, R⁷ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical or —SO₂—R⁹ when R³ is H or a group of the formula II, IIIb or IIIc and, is an aryl or cycloalkyl radical when R³ is a group of the formula IIIa, R⁸ is in each case independently H, halogen or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical, R⁹ is H or an alkyl, alkenyl, alkynyl, aryl or/and heteroaryl radical and n is an integer from 0 to 2, R⁵ is H, an alkyl, alkenyl, alkynyl, carboxyalkyl, carboxyalkenyl, carboxyalkynyl, carboxyaryl or carboxyheteroaryl radical; R² is halogen, C(R⁶)₃, C₂(R⁶)₅, OC(R⁶)₃ or OC²(R⁶)₅, where R⁶ is in each case independently H or halogen and m is an integer from 0 to 4; or salts of said at least one compound, and a pharmaceutically acceptable carrier therefor.
 27. A compound of the formula:

or a salt thereof.
 28. A compound of the formula:

or a salt thereof.
 29. A compound of the formula:

or a salt thereof.
 30. A pharmaceutical composition comprising the compound of claim 27 and a pharmaceutical acceptable carrier therefor.
 31. A pharmaceutical composition comprising the compound of claim 28 and a pharmaceutical acceptable carrier therefor.
 32. A pharmaceutical composition comprising the compound of claim 29 and a pharmaceutical acceptable carrier therefor. 